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Product name

Anti-Tet2 antibody

Description

Rabbit polyclonal to Tet2

Host species

Rabbit

Tested applications

Suitable for: ICC/IF,WB,IP

Species reactivity

Reacts with: Mouse

Immunogen

Synthetic peptide corresponding to Mouse Tet2 aa 1600-1700 conjugated to Keyhole Limpet Haemocyanin (KLH).
Database link: Q4JK59

Positive control

• This antibody gave a positive signal in the following Mouse whole cell lysates: E14Tg2A ; Wild Type Mouse ES Cell Lysate (Tet2 Positive Control)

Form

Liquid

Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.

Storage buffer

pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS

Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.

Purity

Immunogen affinity purified

Clonality

Polyclonal

Isotype

IgG

All lanes : Anti-Tet2 antibody (ab124297) at 1/250 dilution (Milk blocking - 1%)

Lane 1 : E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate
Lane 2 : WT Mouse ES Cell Lysate (Positive Control for Tet2)
Lane 3 : Tet2 Knockout Mouse ES Cell Lysate (Negative Control)

Lysates/proteins at 25 μg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 223 kDa
Observed band size: 223 kDa
Additional bands at: 48 kDa (possible non-specific binding), 65 kDa (possible non-specific binding), 70 kDa (possible non-specific binding)


Exposure time: 4 minutes

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 1% Milk before being incubated with ab124297 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

 

Secondary antibody - goat anti-rabbit HRP (ab97051)

All lanes : Anti-Tet2 antibody (ab124297) at 1/1000 dilution (Incubated for 12 hours at 4°C)

Lane 1 : Wild type mouse embryonic stem cells with Milk, 1 hour at 21°C
Lane 2 : Tet2 KO clone#1 mouse embryonic stem cells with Milk, 1 hour at 21°C
Lane 3 : Tet2 KO clone#2 mouse embryonic stem cells with Milk, 1 hour at 21°C

Lysates/proteins at 20 μg per lane.

Blocking peptides at 5 % per lane.

Secondary
All lanes : HRP conjugated Goat anti-rabbit IgG at 1/10000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 223 kDa


Exposure time: 5 minutes

ICC/IF image of Anti-Tet2 antibody (ab124297) stained WT and Tet2^-/- mouse ES cells. The cells were fixed in PFA, permeabilized using 0.25% Triton X-100, and blocked with 10% BSA for 30 minutes. The cells were then incubated with ab124297 at a 1/400 dilution for 13 hours and 20 minutes at 4^oC. The secondary antibody was a Rhodamine Red-X AffiniPure Donkey anti-Rabbit used at a 1/500 dilution.

All lanes : Anti-Tet2 antibody (ab124297) at 1/500 dilution

Lane 1 : Wild-type embryonic stem cells
Lane 2 : Tet2-/- embryonic stem cells

Lysates/proteins at 20 μg per lane.

Secondary
All lanes : Goat anti-Rabbit (whole molecules) IgG HRP at 1/3000 dilution

Developed using the ECL technique.

Performed under reducing conditions.

Predicted band size: 223 kDa
Observed band size: 250 kDa

Exposure time: 2 minutes

ICC/IF image of Anti-Tet2 antibody (ab124297) stained D3 mouse ES cells. The cells were fixed in 4% PFA, permeabilized using 0.1% Triton X-100, and blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes. The cells were then incubated with ab124297 at a 1/100 dilution for 2 hours at RT. The secondary antibody was a Goat polyclonal Secondary Antibody to Rabbit IgG – H&L Alexa Fluor 488 (ab150077) used at a 1/200 dilution.

 

Secondary antibody - goat anti-rabbit Alexa  488 (ab150077)

Immunoprecipitation of Tet2, using WT and Tet2 KO mouse ES cells. The protein was immunoprecipitated using 300μg whole cell extract, and 3μg of Rabbit polyclonal to Tet2 (ab124297). The same Tet2 antibody was used for the western blot. Tet2 KO mouse ES cells were used as a negative control.

Band: 223kDa: Tet2.

All lanes : Anti-Tet2 antibody (ab124297) at 0.1 mg/ml

Lane 1 : Supernatant from WT mouse embryonic stem cells.
Lane 2 : Supernatant from Tet2 KO mouse embryonic stem cells.
Lane 3 : IP with WT mouse embryonic stem cells.
Lane 4 : IP with Tet2 KO mouse embryonic stem cells.

Lysates/proteins at 300 μg per lane.

Additional bands at: 223 kDa. We are unsure as to the identity of these extra bands.